Protease Composition in Tissue Extracts of Hydrobionts from Antarctic Region: Recent Study | Chapter 13 | Current Research Trends in Biological Science Vol. 1
Aims: Marine hydrobionts, which
grow in extreme conditions, e.g. low temperatures, are an important source of
enzymes with unique properties. By this reason the proteases from cold-water
organisms could have a considerable biotechnological and therefore, commercial
significance. The objective of the current study was to investigate the
proteolytic potential of marine hydrobionts from Antarctic region (an example
of Odontaster validus and Glyptonotus antarcticus). Methodology: SDS-PAGE was carried out for the
determination of protein composition in extracts. The proteolytic activity was
monitored by zymographic technique. Further, the samples were preincubated with
protease inhibitors EDTA, PMSF and SBTI and then total proteolytic (with casein
as substrate) activity was measured. Gel filtration chromatography was applied
for the fractionation of tested extracts. Collagenolytic and trypsin-like
(amidase activity) activities were assessed with help of native collagen type I
and L-BApNA respectively. Results: The results of gelatin zymography provided
evidence for the presence of active enzymes in extracts of both hydrobionts
whereas fibrinogen zymography revealed the presence only one clear area in
extract of O. validus. Specific protease inhibitors were used to identify the
nature of proteases present in tissue of investigated hydrobionts. Based on
this analysis, the proteolytic enzymes in extract of O. validus might be
classified as metal-dependent proteases, whereas the enzymes in extract of G.
antarcticus were most likely trypsin-like proteases. Tissue extracts were
separated by gel filtration chromatography on seven fractions for O. validus
and six fractions for G. antarcticus. Further enzymatic activity assay in
obtained fractions revealed that both hydrobionts possessed significant
collagenolytic activity, which was detected in the first four fractions. Conclusion: The current study gives some
information about protease composition in tissue extracts of hydrobionts of
Antarctic region. It could be useful for better understanding of functional and
catalytic characteristics of proteases from cold-water organisms.
Author (s) Details
Nataliia Raksha
ESC “Institute of Biology and Medicine” of Taras Shevchenko National University of Kyiv, 64/13, Volodymyrska Str., Kyiv 01601, Ukraine.
Tetiana Halenova
ESC “Institute of Biology and Medicine” of Taras Shevchenko National University of Kyiv, 64/13, Volodymyrska Str., Kyiv 01601, Ukraine.
Tetyana Vovk
ESC “Institute of Biology and Medicine” of Taras Shevchenko National University of Kyiv, 64/13, Volodymyrska Str., Kyiv 01601, Ukraine.
Oleksii Savchuk
ESC “Institute of Biology and Medicine” of Taras
Shevchenko National University of Kyiv, 64/13, Volodymyrska Str., Kyiv 01601,
Ukraine.Author (s) Details
Nataliia Raksha
ESC “Institute of Biology and Medicine” of Taras Shevchenko National University of Kyiv, 64/13, Volodymyrska Str., Kyiv 01601, Ukraine.
Tetiana Halenova
ESC “Institute of Biology and Medicine” of Taras Shevchenko National University of Kyiv, 64/13, Volodymyrska Str., Kyiv 01601, Ukraine.
Tetyana Vovk
ESC “Institute of Biology and Medicine” of Taras Shevchenko National University of Kyiv, 64/13, Volodymyrska Str., Kyiv 01601, Ukraine.
Oleksii Savchuk
Lydmila Ostapchenko
ESC “Institute of Biology and Medicine” of Taras Shevchenko National University of Kyiv, 64/13, Volodymyrska Str., Kyiv 01601, Ukraine.
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